Acetic acid

Ceramide/PLGA nanoparticles were fabricated following the previous research30 (link): 25 mg poly lactic-co-glycolic acid (PLGA, Sigma-Aldrich, MO, USA) and 10 mg Ceramide (Doosan, South Korea) were dissolved in 0.5 ml chloroform (Sigma-Aldrich, MO, USA), then mixed with 10 ml 2.5% (w/v) polyvinyl alcohol (Junsei, Japan). The resultant sample was subjected to sonication (Sonictopia, South Korea) for 30 s, after which the chloroform was removed by stirring at 4 °C for 24 h. Next, 0.5 ml of 10 mg/ml chitosan (Sigma-Aldrich, MO, USA) dissolved in 1% acetic acid (Daejung, South Korea) was added to each sample and allowed to react for 2 h to develop a chitosan coating on the nanoparticle.

SO and TE powder were obtained from Ottogi Corp., Ltd. (Anyang, Republic of Korea). XG with a viscosity ranging from 1200 to 1600 cPs in a 1% solution in 1% KCl was purchased from Deosen Biochemical (Ordos) Ltd. (Ordos, Inner Mongolia, China). SL was acquired from Solae LLC. (St. Louis, MO, USA). ρ-Anisidine was purchased from Tokyo Chemical Industry Co., Ltd. (Nihonbashi-honcho, Chuo-ku, Tokyo, Japan). Sodium thiosulfate, Nile red, and Calcofluor white dyes were purchased from Sigma-Aldrich (St. Louis, MO, USA). Isooctane, acetic acid, and potassium iodide (KI) were obtained from Daejung Chemicals (Seoul, Republic of Korea). Commercial butter (Anchor butter) was purchased from Fonterra Cooperative Group Ltd. (Auckland, New Zealand). Whole egg liquid and non-fat dry milk were purchased from KC Feed Co., Ltd. (Yeongcheon, Republic of Korea) and Seoul Milk Co. (Seoul, Republic of Korea), respectively. Sugar and flour were purchased from CJ CheilJedang Co., Ltd. (Seoul, Republic of Korea).

Barium chloride dihydrate, ethanol, iron (II) sulfate heptahydrate, acetic acid, hydrochloric acid, nitric acid, ammonium sulfate, sodium hydroxide, phenol, and sulfuric acid were purchased from Daejung (Seoul, Korea). From Sigma-Aldrich (St. Louis, MO, USA), 2,2-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, 2,2-azobis (2-methylpropionamidine) dihydrochloride, 1,1-diphenyl-2-picrylhydrazyl (DPPH), gallic acid, glucose, Folin and Ciocalteu’s phenol reagent, peroxidase, α-(4-pyridyl N-oxide)-N-tert-butylnitrone, 5,5-dimethyl-1-pyrroline-N-oxide, 2′,7′-dichlorofluorescein diacetate (DCF-DA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), and gum arabic were purchased. RPMI-1640 medium, fetal bovine serum, penicillin-streptomycin, and trypsin were purchased from Gibco (Mississauga, ON, Canada). Sodium carbonate anhydrous (Yakuri, Japan), bicinchoninic acid protein assay kit (Thermo Fisher Scientific, Pittsburgh, PA, USA), bovine serum albumin (Bovogen, East Keilor VIC, Australia) and hydrogen peroxide (Junsei, Tokyo, Japan) were used. Commercial food-grade enzymes (alcalase (Al), α-chymotrypsin (α-chy), flavourzyme (Fla), kojizyme (Koj), neutrase (Neu), papain (Pap), pepsin (Pep), protamex (Pro), and trypsin (Try)) were purchased from Novozyme (Bagsvaerd, Copenhagen, Denmark).

Leucine aminopeptidase (EC 3.4.1.2) microsomal from porcine kidney (LAP), β-glucuronidase (EC 3.2.1.31) from helix pomatia type H-2 (GLU), N-(tert-Butoxycarbonyl)glycine, glycine, triglycine, σ-pinene, triphosgene, triethylamine, N-carboxyanhydride (NCA), 2-aminoethyl methacrylate hydrochloride (2-AMEA), allylamine, 4-nitrophenyl- β-D-glucuronide (PNPG), hydroxybenzotriazole (HOBt), N,N’-diisopropylcarboiimide (DIC), acrylic acid N-hydroxysuccinimide (Ac–NHS ester), deuterium oxide (D₂O), dimethylsulfoxide-d₆ (DMSO-d), chloroform-d (CDCl₃), 2,2-dihydroxyindane-1,3-dione (Ninhydrin) and tris(hydroxymethyl)aminomethane hydrochloride (Tris–HCl) was purchased from Sigma Aldrich (St. Louis, MO, USA). GGRSK was custom-synthesized from Lifetein (Somerset, NJ, USA) and Cosmogenetech (Seoul, South Korea). Urokinase (EC 3.4.4.21)-type plasminogen activator (uPA) was purchased from Fortunachem (Wuhan, China). Acrylic acid (AAc), sodium hydroxide (NaOH), ethyl acetate anhydrous (EA), dimethylformamide (DMF), tetrahydrofuran (THF), chloroform (CCl₄), methanol (MeOH), n-butanol (BuOH), acetic acid, acetonitrile, and trifluoroacetic acid were purchased from Daejung (South Korea). 2-hydroxy-2-methlypropiophenone (Darocur 1173) and dimethylsulfoxide anhydrous (DMSO) were purchased from TCI (Tokyo, Japan).

Chitosan was sourced from Sigma–Aldrich Gmbh Darmstadt, Germany and acetic acid was purchased from DAEJUNG, Gyeonggi-do, South Korea. PEO (Mw 600KD) was purchased from Polysciences, Inc., Niles, IL, USA. Clove essential oil (CEO) was purchased from Mueller Hinton broth Sigma–Aldrich Gmbh, Darmstadt, Germany. All chemicals were used without any further purification.

Chitosan (DDA: 75–85%; Mw: 50–190 kDa), ferric chloride hexahydrate (FeCl₃·6H₂O), poly(vinyl alcohol) (PVA), sodium hydroxide (NaOH), α-glucosidase (Saccharomyces cerevisiae), α-amylase (porcine pancreas), acarbose, 4-nitrophenyl α-D-glucopyranoside (PNPG), antioxidant (DPPH and ABTS), and starch were procured from Sigma-Aldrich, Republic of Korea. Cell viability assay kit (Cellomax™) was purchased from MediFab, Republic of Korea. L7012 LIVE/DEAD BacLight Bacterial viability kits from ThermoFisher. Acetic acid was procured from the Daejung chemicals & metals co., Ltd., Siheung-si, South Korea. The manufacturer’s information of all the fluorescent staining and culture medium used in the present study is mentioned in our earlier paper. Pinus densiflora leaves were collected from Kangwon National University campus, Chuncheon, Republic of Korea.